• Journal of Food Hygiene and Safety
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• v.23 no.2
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• pp.121-128
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• 2008

The aim of this study was to develop a LightCycler-based real time PCR (LC-PCR) assay and to evaluate its diagnostic use for the detection of Staphylococcus aureus in raw milk samples. Following amplification of 113 bp of coa gene encoding an coagulase precursor specific for Staphylococcus aureus, melting curve and DNA sequencing analysis was performed to verify the specificity of the PCR products. Amplification of 209 bp gene encoding an altered penicillin-binding protein, PBP2a (mecA), melting curve analysis and DNA sequencing analysis was performed to verify methicillin resistance Staphylococcus aureus (MRSA). According to this study, 6 of 647 raw milk samples showed S. aureus positive and 2 of them showed a mecA positive and the detection limit was 10 fg of DNA. And we also isolated Staphylococcus chromogenes a causative agent of exudative epidermitis in pigs and cattle from 3 samples.

• Journal of Food Hygiene and Safety
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• v.26 no.2
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• pp.114-119
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• 2011

In this study, two duplex real-time PCR approach with melting curve analysis is presented for the detection of Escherichia coli O157:H7, Listeria monocytogenes, Salmonella spp. and Staphylococcus aureus, which are important food-borne bacterial pathogens usually present in fresh and/or minimally processed vegetables. Reaction conditions were adjusted for the simultaneous amplification and detection of specific fragments in the ${\beta}$-glucuronidase (uidA, E. coli), thermonuclease (nuc, S. aureus), hemolycin (hly, L. monocytogenes) and tetrathionate reductase (ttr, Salmonella spp.) genes. Melting curve analysis using a SYBR Green I real-time PCR approach showed characteristic $T_m$ values demonstrating the specific and efficient amplification of the four pathogens; $80.6{\pm}0.9^{\circ}C$, $86.9{\pm}0.5^{\circ}C$, $80.4{\pm}0.6^{\circ}C$ and $88.1{\pm}0.11^{\circ}C$ for S. aureus, E. coli O157:H7, L. monocytogenes and Salmonella spp., respectively. For all the pathogens, the two duplex, real-time PCR was equally sensitive to uniplex real-time PCR, using same amounts of purified DNA, and allowed detection of 10 genome equivalents. When our established duplex real-time PCR assay was applied to artificially inoculated fresh lettuce, the detection limit was $10^3$ CFU/g for each of these pathogens without enrichment. The results from this study showed that the developed duplex real-time PCR with melting curve analysis is promising as a rapid and cost-effective test method for improving food safety.

• Journal of Crop Science and Biotechnology
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• v.11 no.2
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• pp.91-100
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• 2008

Melting curve analysis of fluorescently labeled DNA fragments is used extensively for genotyping single nucleotide polymorphism(SNP). Here, we evaluated a SNP genotyping method by melting curve analysis with the two probe chemistries in a 384-well plate format on a Roche LightCycler 480. The HybProbe chemistry is based on the fluorescence resonance energy transfer(FRET) and the SimpleProbe chemistry uses a terminal self-quenching fluorophore. We evaluated FRET HybProbes and SimpleProbes for two SNP sites closely linked to two quantitative trait loci(QTL) for southern root-knot nematode resistance. These probes were used to genotype the two parents and 94 $F_2$ plants from the cross of PI 96354$\times$Bossier. The SNP genotypes of all samples determined by the LightCycler software agreed with previously determined SSR genotypes and the SNP genotypes determined on a Luminex 100 flow cytometry instrument. Multiplexed HybProbes for the two SNPs showed a 98.4% success rate and 100% concordance between repeats two of the same 96 DNA samples. Also, we developed a HybProbe assay for the Rcs3 gene conditioning broad resistance to the frogeye leaf spot(FLS) disease. The LightCycler 480 provides rapid PCR on 384-well plate and allows simultaneous amplification and analysis in approximately 2 hours without any additional steps after amplification. This allowed for a reduction of the potential contamination of PCR products, simplicity, and enablement of a streamlined workflow. The melting curve analysis on the LightCycler 480 provided high-throughput and rapid SNP genotyping and appears highly effective for marker-assisted selection in soybean.

• Journal of the Korean Institute of Electrical and Electronic Material Engineers
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• v.31 no.6
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• pp.427-432
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• 2018

High-capacity secondary batteries can cause explosion hazards owing to microcurrent variations or current surges that occur in short circuits. Consequently, complete safety cannot be achieved with general protection that is limited to a mere current fuse. Hence, in the case of secondary batteries, it is necessary for the protector to limit the inrush current in a short circuit, and to detect the current during microcurrent variations. To serve this purpose, a fuse can be employed for the secondary battery protection circuit with current detection. This study aims at designing a protection device that can stably operate in the hazardous circumstances associated with high-capacity secondary batteries. To achieve the said objective, a detecting fuse was designed from an alloy of low melting point elements for securing stability in abnormal current states. Experimental results show that the operating I-T and V-T characteristic constraints can be satisfied by employing the proposed current detecting self-contained low melting point fuse, and through the resistance of the heating resistor. These results thus verify that the proposed protection device can prevent the hazards of short circuit current surges and microcurrent variations of secondary batteries.

• Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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• 1996.11a
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• pp.214-217
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• 1996

In this paper. we deal wish the I-t characteristic of law voltage distribution fuse. It is used to be thermal characteristic in being produced at fuse element part. The elements are divided low temperature melting element(LTME) by high temperature melting element(HTME). Those parts make of coordination. The characteristic of fuse is decided by material and design etc. used at element. We analysis I-t characteristic curve by using the numerical method. And we compared the curve of simulation with that of experiment

• The Transactions of The Korean Institute of Electrical Engineers
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• v.67 no.12
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• pp.1619-1625
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• 2018

Portable electronic devices secondary batteries can cause fire and explosion due to micro-current change in addition to the situation of short-circuit inrush current, safety can not be secured with a general operation limited current fuse. Therefore, in secondary battery, it is necessary for the protector to satisfy both the limit current type operation in the open-short-circuit inrush current and the current detection operation characteristic in the micro current change situation and for this operation, a fuse for the current detection type secondary battery protection circuit can be applied. The purpose of this study is to design a protection device that operates stably in the hazardous situation of small capacity secondary battery for portable electronic devices through the design of low melting fuse elements alloy of sensing type fuse and secures stability in abnormal current state. As a result of the experiment, I-T and V-T operation characteristics are satisfied in a the design of the alloy of the current sensing type self-contained low melting point fuse and the resistance of the heating resistor. It is confirmed that it can prevent accidents of short circuit over-current and micro current change of secondary battery.

• Bulletin of the Korean Chemical Society
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• v.18 no.1
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• pp.38-43
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• 1997

Poly(ethylene 2, 6-naphthalate-co-tetramethylene 2, 6-naphthalate) (PEN/PBN) copolymers were synthesized and studied by 13C NMR spectroscopy, DSC analysis and X-ray diffraction. A minimum in the melting point vs. composition curve was found at approximately 60 mol% tetramethylene 2, 6-naphthalate. The PEN/PBN copolymers were shown to be statistically random throughout the range of 1, 4-butanediol compositions. The melting point depression behavior of annealed PEN/PBN copolymers depended upon the sequence propagation probability, PS, which is suggested by indivisual crystal formation of two pure comonomers; that is, ethylene-naphthalate-ethylene, EE, and tetramethylene-naphthalate-tetramethylene, BB. However, it can be seen from the X-ray curve that the peaks of PEN/PBN copolymers appear from a crystal lattice which is governed only by the rich component between two different aliphatic units in the copolymer composition.

• Proceedings of the Korea Institute of Applied Superconductivity and Cryogenics Conference
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• 1999.02a
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• pp.161-164
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• 1999

$^{3}$He melting curve thermometer is made, tested, and compared with Greywall scale. The experimental results show that we can identify the temperature within a few percentage. To improve the accuracy of our thermometer we need temperature fixed points like $^{3}$He superfluid phase transition temperature.

• Journal of Sensor Science and Technology
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• v.18 no.2
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• pp.116-121
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• 2009

Pressure-controlled sodium heat pipe furnace was designed and fabricated, and its characteristics was investigated. Pressure control system was controlled within ${\pm}0.5\;Pa$ at 150 kPa and the stability of pressure was decreased to ${\pm}2.5\;Pa$, when the pressure-controlled system connected with the heat pipe. The melting curve of Ag fixed-point realized well by the adiabatic method using the pressure-controlled sodium heat pipe furnace and its accuracy showed ${\pm}2.27\;mK$ from the calculation of 20% to 80% at the plateau. The freezing curve of Ag fixed-point also realized and its plateau value was 2.23 mK lower than that of the melting curve.

• Parasites, Hosts and Diseases
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• v.55 no.6
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• pp.601-606
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• 2017

Cystoisospora is responsible for morbidity in immunocompromised patients. PCR is sensitive for diagnosing Cystoisospora; however, it needs reevaluation for differential molecular diagnosis of cystoisosporiasis. We aimed at evaluating melting curve analysis (MCA) after real-time PCR (qPCR) in diagnosis and genotyping of Cystoisospora as an alternative to conventional PCR. We included 293 diarrheic stool samples of patients attending the Department of Clinical Oncology and Nuclear Medicine of Cairo University Hospitals, Egypt. Samples were subjected to microscopy, nested PCR (nPCR), and qPCR targeting the internal transcribed spacer 2 region (ITS2) of the ribosomal RNA (r RNA) gene followed by melting temperatures ($T_ms$) analysis and comparing the results to PCR-RFLP banding patterns. Using microscopy and ITS2-nPCR, 3.1% and 5.8% of cases were Cystoisospora positive, respectively, while 10.9% were positive using qPCR. Genotyping of Cystoisospora by qPCR-MCA revealed 2 genotypes. These genotypes matched with 2 distinct melting peaks with specified $T_ms$ at $85.8^{\circ}C$ and $88.6^{\circ}C$, which indicated genetic variation among Cystoisospora isolates in Egypt. Genotype II proved to be more prevalent (65.6%). HIV-related Kaposi sarcoma and leukemic patients harbored both genotypes with a tendency to genotype II. Genotype I was more prevalent in lymphomas and mammary gland tumors while colorectal and hepatocellular tumors harbored genotype II suggesting that this genotype might be responsible for the development of cystoisosporiasis in immunocompromised patients. Direct reliable identification and differentiation of Cystoisospora species could be established using $qPCR-T_ms$ analysis which is useful for rapid detection and screening of Cystoisospora genotypes principally in high risk groups.